Compounds specific for the human α1d adrenergic receptor and uses thereof

ABSTRACT

This invention is directed towards a method of inhibiting activation of a human α 1d  adrenergic receptor which comprises contacting the receptor with a compound so as to inhibit activation of the receptor, wherein the compound binds selectively to a human α 1d  adrenergic receptor. This invention provides for a compound which binds selectively to a human α 1d  adrenergic receptor. The invention further provides a pharmaceutical composition comprising a therapeutically effective amount of the above-defined compounds and a pharmaceutically acceptable carrier. This invention further provides for a method of treating a subject afflicted with a disease which is susceptible to treatment by antagonism of the human α 1d  adrenergic receptor which comprises administering to the subject an amount of the above defined compounds effective to treat the disease.

This application is a continuation of PCT International Application No.PCT/US99/16101, filed Jul. 16, 1999, designating the United States ofAmerica, which is a continuation-in-part of U.S. Ser. No. 09/118,323,filed Jul. 17, 1998 now abandoned, the contents of which are herebyincorporated by reference into the present application.

The invention disclosed herein was made with Government support underSmall Business Innovation Research (SBIR) Program Grant No. 2 R44NS33418-02 from the National Institute of Neurological Disorders andStroke, National Institutes of Health. Accordingly, the U.S. Governmenthas certain rights in the invention.

Throughout this application, various references are referred to withinparentheses. Disclosures of these publications in their entireties arehereby incorporated by reference into this application to more fullydescribe the state of the art to which this invention pertains. Fullbibliographic citation for these references may be found at the end ofthis application, preceding the sequence listings and the claims.

BACKGROUND OF THE INVENTION

The designation “α_(1d)” is the appellation recently approved by theIUPHAR Nomenclature Committee for the previously designated “α_(1a)”cloned subtype as outlined in the Pharmacological Reviews (Hieble, etal., 1995). The designation “α_(1d)” is used throughout this applicationand the supporting tables and figures to refer to this receptor subtype.At the same time, the receptor formerly designated “α_(1c)” was renamed“α_(1a)”. The new nomenclature is used throughout this application.Stable cell lines expressing these receptors are described herein;however, these cell lines were deposited with the American Type CultureCollection (ATCC) under the old nomenclature (infra). In each case,lowercase letters are used to designate cloned receptors (i.e., α_(1a),α_(1b), α_(1d)) and uppercase letters are used to designatepharmacologically defined native receptors (i.e., α_(1A), α_(1B), andα_(1D))

α-Adrenergic receptors (McGrath et al., 1989) are specific neuroreceptorproteins both located in the peripheral and central nervous systems andin tissues and organs throughout the body. These receptors are importantswitches for controlling many physiological functions and, thus,represent important targets for drug development. In fact, manyα-adrenergic drugs have been developed over the past 40 years. Examplesinclude clonidine, phenoxybenzamine and prazosin (for treatment ofhypertension), naphazoline (a nasal decongestant), and apraclonidine(for treatment of glaucoma). α-Adrenergic drugs can be dividedfunctionally into two distinct classes: agonists (e.g., clonidine andnaphazoline), which mimic the receptor activation properties of theendogenous neurotransmitter norepinephrine, and antagonists (e.g.,phenoxybenzamine and prazosin), which act to block the effects ofnorepinephrine. Many of these drugs are effective, but also produceunwanted side effects (e.g., clonidine produces dry mouth and sedationin addition to its antihypertensive effects).

During the past 15 years, a more precise understanding of α-adrenergicreceptors and their drugs has evolved through increased scientificscrutiny. Prior to 1977, only one α-adrenergic receptor was known toexist. Between 1977 and 1988, it was accepted by the scientificcommunity that at least two α-adrenergic receptor types—α₁ andα₂—existed in the central and peripheral nervous systems. Since 1988,new techniques in molecular biology have led to the identification of atleast six α-adrenergic receptors which exist throughout the central andperipheral nervous systems: α_(1A) (new nomenclature), α_(1B), α_(1D)(new nomenclature), α_(2A), α_(2B) and α_(2C) (Bylund, D. B., 1992). Inmany cases, it is not known precisely which physiological responses inthe body are controlled by each of these receptors. In addition, currentα-adrenergic drugs are not selective for any particular α-adrenergicreceptor. Many of these drugs produce untoward side effects that may beattributed to their poor α-adrenergic receptor subtype selectivity.

This invention is directed to compounds which are selective antagonistsfor cloned human α_(1d) receptors. This invention is also related to theuse of these compounds as antihypertensive agents (Deng, F. X. et al.,1996). Experimental evidence presented herein indicates that thesecompounds, while effective at reducing blood pressure in hypertensiveindividuals, will be devoid of hypotensive actions in normotensiveindividuals.

This invention is also related to the use of these compounds for thetreatment of Raynaud's disease and for treating bladder instabilityassociated with urinary incontinence (Broten, et al., 1998).

SUMMARY OF THE INVENTION

The invention is directed to a method of inhibiting activation of ahuman α_(1d) adrenergic receptor which comprises contacting the receptorwith a compound so as to inhibit activation of the receptor, wherein thecompound binds to the human α_(1d) adrenergic receptor with a bindingaffinity which is at least ten-fold higher than the binding affinitywith which the compound binds to (i) a human α_(1a) adrenergic receptorand (ii) a human α_(1b) adrenergic receptor, and the compound binds tothe human α_(1d) adrenergic receptor with a binding affinity which isgreater than the binding affinity with which the compound binds to ahuman 5-HT_(1a) receptor.

This invention is additionally directed to a method of inhibitingactivation of a human α_(1d) adrenergic receptor which comprisescontacting the receptor with a compound so as to inhibit activation ofthe receptor, wherein the compound has the structure:

wherein m is an integer from 0 to 2; wherein n is an integer from 0 to2;

wherein Y is

wherein Z is

wherein R1 and R2 (i) are independently H, branched or unbranched C₁-C₆alkyl or alkoxy, branched or unbranched C₂-C₆ alkenyl or alkynyl,branched or unbranched C₁-C₆ hydroxyalkyl, hydroxy, substituted orunsubstituted aryl or aryl-(C₁-C₆)-alkyl, or substituted orunsubstituted heteroaryl or heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, hydroxy, branched orunbranched C₁-C₆ alkyl or alkoxy group, or branched or unbranched C₂-C₆alkenyl or alkynyl group; or (ii) taken together form a substituted orunsubstituted cycloalkyl ring containing 3-10 carbons, wherein thesubstituent if present is a branched or unbranched C₁-C₆ alkyl group orbranched or unbranched C₂-C₆ alkenyl or alkynyl group;

wherein R3 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R4 is H or CH₃;

wherein R5 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R6 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₃-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, C₀ ₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R7 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, aryl,aryl-(C₁-C₆)-alkyl, CO₂R14, CON(R14)₂, substituted C₁-C₆ alkyl,substituted aryl, wherein the substituent is N(R14)₂, halogen, OR14 orSR14;

wherein R8 is H or CH₃;

wherein R9 is H, F, Cl, Br, branched or unbranched C₁-C₆ alkyl oralkoxy, CN; wherein R10 is H or F; wherein R11 is H, F, Cl, Br, I, CN,branched or unbranched C₁-C₆ alkyl or alkoxy; wherein R12 is H, F, Cl,CN, branched or unbranched C₁-C₆ alkyl or alkoxy; wherein R13 is H or F;wherein X is N or CH; with the proviso that when R11 and R12 are each H,then R9 is F;

and wherein R14 is independently H or branched or unbranched C₁-C₆alkyl.

This invention is additionally directed to a compound having thestructure:

wherein n is an integer from 0 to 2; wherein m is an integer from 0 to2;

wherein Y is

wherein Z is

wherein R1 and R2 (i) are independently H, branched or unbranched C₁-C₆alkyl or alkoxy, branched or unbranched C₂-C₆, alkenyl or alkynyl,branched or unbranched C₁-C₆ hydroxyalkyl, hydroxy, substituted orunsubstituted aryl or aryl-(C₁-C₆)-alkyl, or substituted orunsubstituted heteroaryl or heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, hydroxy, branched orunbranched C₁-C₆ alkyl or alkoxy group, or branched or unbranched C₂-C₆alkenyl or alkynyl group; or (ii) taken together form a substituted orunsubstituted cycloalkyl ring containing 3-10 carbons, wherein thesubstituent if present is a branched or unbranched C₁-C₆ alkyl group orbranched or unbranched C₂-C₆ alkenyl or alkynyl group;

wherein R3 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R4 is H or CH₃;

wherein R5 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R6 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂ SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R7 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, aryl,aryl-(C₁-C₆)-alkyl, CO₂R14, CON(R14)₂, substituted C₁-C₆ alkyl,substituted aryl, wherein the substituent is N(R14)₂, halogen, OR14 orSR14;

wherein R8 is H or CH₃;

wherein R10 is H or F; wherein R11 is H, F, Cl, Br, I, CN, branched orunbranched C₁-C₆ alkyl or alkoxy; wherein R12 is H, F, Cl, CN, branchedor unbranched C₂-C₆ alkyl or alkoxy; wherein R13 is H or F; wherein X isN or CH; and wherein R14 is independently H or branched or unbranchedC₁-C₆ alkyl.

This invention is additionally directed towards a pharmaceuticalcomposition comprising a therapeutically effective amount of thecompound of this invention and a pharmaceutically acceptable carrier.

This invention is additionally directed towards a pharmaceuticalcomposition obtained by combining a therapeutically effective amount ofa compound of this invention and a pharmaceutically acceptable carrier.

This invention is additionally directed towards a process for making apharmaceutical composition comprising combining a therapeuticallyeffective amount of a compound of this invention and a pharmaceuticallyacceptable carrier.

This invention is additionally directed towards a process of making acompound with structure:

which comprises reacting a compound with structure:

with a compound

to form the compound,

wherein Y is

wherein Z is

wherein R1 and R2 (i) are independently H, branched or unbranched C₁-C₆alkyl or alkoxy, branched or unbranched C₂-C₆ alkenyl or alkynyl,branched or unbranched C₁-C₆ hydroxyalkyl, hydroxy, substituted orunsubstituted aryl or aryl-(C₁-C₆)-alkyl, or substituted orunsubstituted heteroaryl or heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, hydroxy, branched orunbranched C₁-C₆ alkyl or alkoxy group, or branched or unbranched C₂-C₆alkenyl or alkynyl group; or (ii) taken together form a substituted orunsubstituted cycloalkyl ring containing 3-10 carbons, wherein thesubstituent if present is a branched or unbranched C₁-C₆ alkyl group orbranched or unbranched C₂-C₆ alkenyl or alkynyl group;

wherein R3 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R4 is H or CH₃;

wherein R5 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R6 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂ CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂; and wherein R14 is independently H or branched orunbranched C₁-C₆ alkyl.

This invention is additionally directed towards a method of treating asubject afflicted with a disease which is susceptible to treatment byantagonism of the human α_(1d) adrenergic receptor which comprisesadministering to the subject an amount of the compound of this inventioneffective to treat the disease.

This invention is additionally directed towards a method of treating asubject afflicted with hypertension which comprises administering to thesubject an amount of the compound of this invention effective to treathypertension.

This invention is directed towards a method of treating a subjectafflicted with Raynaud's disease which comprises administering to thesubject an amount of the compound of this invention effective to treatRaynaud's disease.

This invention is directed towards a method of treating a subjectafflicted with urinary incontinence which comprises administering to thesubject an amount of the compound of this invention effective to treaturinary incontinence.

This invention is directed towards a method of treating urinaryincontinence in a subject which comprises administering to the subject atherapeutically effective amount of a α_(1d) antagonist which binds tothe human α_(1d) adrenergic receptor with a binding affinity which is atleast ten-fold higher than the binding affinity with which the α_(1d)antagonist binds to (i) a human α_(1a) adrenergic receptor and (ii) ahuman α_(1b) adrenergic receptor, and the α_(1d) antagonist binds to thehuman α_(1d) adrenergic receptor with a binding affinity which isgreater than the binding affinity with which the α_(1d) antagonist bindsto a human 5-HT_(1a) receptor.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A-1C FIGS. 1A-1C show the structures of the compounds describedherein in the Examples.

FIG. 2 Effects of α_(1D) and α_(1A) receptor blockade on contractions ofthe rat caudal artery evoked by electrical stimulation of intrinsicnerves.

FIG. 3 Effects of α_(1D) and α_(1A) receptor blockade on contractions ofthe rat caudal artery evoked by application of norepinephrine to thetissue bath.

FIG. 4 Cardiovascular effects of BMY 7383 in urethane-anesthetized ratsand effect (antagonism) of WAY 100635. n=4; 10 min post-dose.

FIG. 5 Cardiovascular effects of Compound 7 in urethane-anesthetizedrats and effect (antagonism) of WAY 100635. n=6; 10 min post-dose.

FIG. 6 Effect of the selective α_(1D) adrenoceptor antagonist Compound 7on mean arterial blood pressure in conscious, normotensive rats.

DETAILED DESCRIPTION OF THE INVENTION

This invention is directed towards a method of inhibiting activation ofa human α_(1d) adrenergic receptor which comprises contacting thereceptor with a compound so as to inhibit activation of the receptor,wherein the compound binds to the human α_(1d) adrenergic receptor witha binding affinity which is at least ten-fold higher than the bindingaffinity with which the compound binds to (i) a human α_(1a) adrenergicreceptor and (ii) a human α_(1b) adrenergic receptor, and the compoundbinds to the human α_(1d) adrenergic receptor with a binding affinitywhich is greater than the binding affinity with which the compound bindsto a human 5-HT_(1a) receptor.

In one embodiment, the compound binds to the human α_(1d) adrenergicreceptor with a binding affinity which is at least 25-fold higher thanthe binding affinity with which the compound binds to (i) the humanα_(1a) adrenergic receptor and (ii) the human α_(1b) adrenergicreceptor, and the compound binds to the human α_(1d) adrenergic receptorwith a binding affinity which is at least ten-fold higher than thebinding affinity with which the compound binds to the human 5-HT_(1a)receptor.

In another embodiment, the compound binds to the human α_(1d) adrenergicreceptor with a binding affinity which is at least 25-fold higher thanthe binding affinity with which the compound binds to (i) the humanα_(1a) adrenergic receptor, (ii) the human α_(1b) adrenergic receptor,and (iii) the human 5-HT_(1a) receptor.

In another embodiment, the compound binds to the human α_(1d) adrenergicreceptor with a binding affinity which is at least 100-fold higher thanthe binding affinity with which the compound binds to (i) the humanα_(1a) adrenergic receptor, (ii) the human α_(1b) adrenergic receptor,and (iii) the human 5-HT_(1a) receptor.

This invention is additionally directed to a method of inhibitingactivation of a human α_(1d) adrenergic receptor which comprisescontacting the receptor with a compound so as to inhibit activation ofthe receptor, wherein the compound has the structure:

wherein m is an integer from 0 to 2; wherein n is an integer from 0 to2;

wherein Y is

wherein Z is

wherein R1 and R2 (i) are independently H, branched or unbranched C₁-C₆alkyl or alkoxy, branched or unbranched C₂-C₆ alkenyl or alkynyl,branched or unbranched C₁-C₆ hydroxyalkyl, hydroxy, substituted orunsubstituted aryl or aryl-(C₁-C₆)-alkyl, or substituted orunsubstituted heteroaryl or heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, hydroxy, branched orunbranched C₁-C₆ alkyl or alkoxy group, or branched or unbranched C₂-C₆alkenyl or alkynyl group; or (ii) taken together form a substituted orunsubstituted cycloalkyl ring containing 3-10 carbons, wherein thesubstituent if present is a branched or unbranched C₁-C₆ alkyl group orbranched or unbranched C₂-C₆ alkenyl or alkynyl group;

wherein R3 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R4 is H or CH₃;

wherein R5 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R6 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R7 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, aryl,aryl-(C₁-C₆)-alkyl, CO₂R14, CON(R14)₂, substituted C₁-C₆ alkyl,substituted aryl, wherein the substituent is N(R14)₂, halogen, OR14 orSR14;

wherein R8 is H or CH₃;

wherein R9 is H, F, Cl, Br, branched or unbranched C₁-C₆ alkyl oralkoxy, CN; wherein R10 is H or F; wherein R11 is H, F, Cl, Br, I, CN,branched or unbranched C₁-C₆ alkyl or alkoxy; wherein R12 is H, F, Cl,CN, branched or unbranched C₁-C₆ alkyl or alkoxy; wherein R13 is H or F;wherein X is N or CH; with the proviso that when R11 and R12 are each H,then R9 is F;

and wherein R14 is independently H or branched or unbranched C₁-C₆alkyl.

In one embodiment, the compound has the structure:

In another embodiment, the compound has the structure:

In another embodiment, the compound has the structure:

In another embodiment, the compound has the structure:

In another embodiment, the compound has the structure:

In another embodiment, the compound has the structure:

In another embodiment, the compound has the structure:

This invention is additionally directed to a compound having thestructure:

wherein n is an integer from 0 to 2; wherein m is an integer from 0 to2;

wherein Y is

wherein Z is

wherein R1 and R2 (i) are independently H, branched or unbranched C₁-C₆alkyl or alkoxy, branched or unbranched C₂-C₆ alkenyl or alkynyl,branched or unbranched C₁-C₆ hydroxyalkyl, hydroxy, substituted orunsubstituted aryl or aryl-(C₁-C₆)-alkyl, or substituted orunsubstituted heteroaryl or heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, hydroxy, branched orunbranched C₁-C₆ alkyl or alkoxy group, or branched or unbranched C₂-C₆alkenyl or alkynyl group; or (ii) taken together form a substituted orunsubstituted cycloalkyl ring containing 3-10 carbons, wherein thesubstituent if present is a branched or unbranched C₁-C₆ alkyl group orbranched or unbranched C₂-C₆ alkenyl or alkynyl group;

wherein R3 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₆cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R4 is H or CH₃;

wherein R5 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R6 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R7 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, aryl,aryl-(C₁-C₆)-alkyl, CO₂R14, CON(R14)₂, substituted C₁-C₆ alkyl,substituted aryl, wherein the substituent is N(R14)₂, halogen, OR14 orSR14;

wherein R8 is H or CH₃;

wherein R10 is H or F; wherein R11 is H, F, Cl, Br, I, CN, branched orunbranched C₁-C₆ alkyl or alkoxy; wherein R12 is H, F, Cl, CN, branchedor unbranched C₁-C₆ alkyl or alkoxy; wherein R13 is H or F; wherein X isN or CH; and wherein R14 is independently H or branched or unbranchedC₁-C₆ alkyl.

In one embodiment, the compound has the structure:

In another embodiment, the compound has the structure:

In another embodiment, the compound has the structure:

In another embodiment, the compound has the structure:

In another embodiment, the compound has the structure:

In another embodiment, the compound has the structure:

The compounds of the present invention may be present as singleenantiomers, diasteriomers, or cis or trans isomers; or two or more ofthe compounds may be present to form a mixture of enantiomers,diasteriomers, or isomers, including a racemic mixture.

The invention also provides for the (−) and (+) enantiomers of allcompounds of the subject application described herein.

The invention further provides for the cis, trans, erythro, and threoisomers of all of the compounds of the subject application describedherein. It is noted herein that the terms “cis”, “trans”, “erythro”, and“threo” correspond to relative stereochemistry, as determined, forexample, by NOE (Nuclear Overhauser Effect) experiments.

The compounds of the present invention are preferably at least 80% pure,more preferably at least 90% pure, and most preferably at least 95%pure.

In the present invention, the term aryl is used to include phenyl,benzyl, benzoyl, or naphthyl, and the term heteroaryl is used to includepyrazinyl, pyrrolyl, furanyl, thiophenyl, pyridyl, imidazolyl,imidazolinyl, indolyl, benzimidazolyl, benzfuranyl, pyrimidyl,benzothiophenyl, isoquinolyl, or quinolyl. The term aryl-(C₁-C₆)-alkylis used to designate an C₁-C₆ alkyl chain substituted with an aryl groupand the term heteroaryl-(C₁-C₆)-alkyl is used to designate a C₁-C₆ alkylchain substituted with a heteroaryl group.

Abbreviations used in the specification, in particular the Schemes andExamples, are as follows:

TFA trifluoroacetic acid HCl hydrochloric acid C₅H₅N pyridine SOCl₂thionyl chloride LiAlH₄ lithium aluminum hydride BuOH butyl alcoholNa₂CO₃ sodium carbonate NH₄Cl ammonium chloride H₂ hydrogen Pd/Cpalladium on charcoal Pd palladium dba dibenzylideneacetone BINAP2,2′-bis(diphenylphosphino)-1,1′- binaphthyl

The invention is additionally directed towards a pharmaceuticalcomposition comprising a therapeutically effective amount of thecompound of the invention and a pharmaceutically acceptable carrier.

In one embodiment, the amount of the compound is an amount from about0.01 mg to about 800 mg.

In another embodiment, the amount of the compound is from about 0.1 mgto about 300 mg.

In another embodiment, the amount of the compound is from about 1 mg toabout 20 mg.

In another embodiment, the carrier is a liquid.

In another embodiment, the carrier is a solid.

In another embodiment, the carrier is a gel.

The invention is additionally directed towards a pharmaceuticalcomposition obtained by combining a therapeutically effective amount ofa compound of the invention and a pharmaceutically acceptable carrier.

The invention is additionally directed towards a process for making apharmaceutical composition comprising combining a therapeuticallyeffective amount of a compound of the invention and a pharmaceuticallyacceptable carrier.

In the subject invention, a “therapeutically effective amount” is anyamount of a compound which, when administered to a subject sufferingfrom a disease against which the compounds are effective, causesreduction, remission, or regression of the disease or reduction of orrelief from symptoms of the disease. In the practice of this invention,the “pharmaceutically acceptable carrier” is any physiological carrierknown to those of ordinary skill in the art useful in formulatingpharmaceutical compositions.

In one embodiment the pharmaceutical carrier may be a liquid and thepharmaceutical composition would be in the form of a solution. Inanother embodiment, the pharmaceutically acceptable carrier is a solidand the composition is in the form of a powder or tablet. In a furtherembodiment, the pharmaceutical carrier is a gel and the composition isin the form of a suppository or cream. In a further embodiment thecompound may be formulated as a part of a pharmaceutically acceptabletransdermal patch.

A solid carrier can include one or more substances which may also act asflavoring agents, lubricants, solubilizers, suspending agents, fillers,glidants, compression aids, binders or tablet-disintegrating agents; itcan also be an encapsulating material. In powders, the carrier is afinely divided solid which is in admixture with the finely dividedactive ingredient. In tablets, the active ingredient is mixed with acarrier having the necessary compression properties in suitableproportions and compacted in the shape and size desired. The powders andtablets preferably contain up to 99% of the active ingredient. Suitablesolid carriers include, for example, calcium phosphate, magnesiumstearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose,polyvinylpyrrolidine, low melting waxes and ion exchange resins.

Liquid carriers are used in preparing solutions, suspensions, emulsions,syrups, elixirs and pressurized compositions. The active ingredient canbe dissolved or suspended in a pharmaceutically acceptable liquidcarrier such as water, an organic solvent, a mixture of both orpharmaceutically acceptable oils or fats. The liquid carrier can containother suitable pharmaceutical additives such as solubilizers,emulsifiers, buffers, preservatives, sweeteners, flavoring agents,suspending agents, thickening agents, colors, viscosity regulators,stabilizers or osmo-regulators. Suitable examples of liquid carriers fororal and parenteral administration include water (partially containingadditives as above, e.g. cellulose derivatives, preferably sodiumcarboxymethyl cellulose solution), alcohols (including monohydricalcohols and-polyhydric alcohols, e.g. glycols) and their derivatives,and oils (e.g. fractionated coconut oil and arachis oil). For parenteraladministration, the carrier can also be an oily ester such as ethyloleate and isopropyl myristate. Sterile liquid carriers are useful insterile liquid form compositions for parenteral administration. Theliquid carrier for pressurized compositions can be halogenatedhydrocarbon or other pharmaceutically acceptable propellent.

Liquid pharmaceutical compositions which are sterile solutions orsuspensions can be utilized by for example, intramuscular, intrathecal,epidural, intraperitoneal or subcutaneous injection. Sterile solutionscan also be administered intravenously. The compounds may be prepared asa sterile solid composition which may be dissolved or suspended at thetime of administration using sterile water, saline, or other appropriatesterile injectable medium. Carriers are intended to include necessaryand inert binders, suspending agents, lubricants, flavorants,sweeteners, preservatives, dyes, and coatings.

The compound can be administered orally in the form of a sterilesolution or suspension containing other solutes or suspending agents,for example, enough saline or glucose to make the solution isotonic,bile salts, acacia, gelatin, sorbitan monoleate, polysorbate 80 (oleateesters of sorbitol and its anhydrides copolymerized with ethylene oxide)and the like.

The compound can also be administered orally either in liquid or solidcomposition form. Compositions suitable for oral administration includesolid forms, such as pills, capsules, granules, tablets, and powders,and liquid forms, such as solutions, syrups, elixirs, and suspensions.Forms useful for parenteral administration include sterile solutions,emulsions, and suspensions.

Optimal dosages to be administered may be determined by those skilled inthe art, and will vary with the particular compound in use, the strengthof the preparation, the mode of administration, and the advancement ofthe disease condition. Additional factors depending on the particularsubject being treated will result in a need to adjust dosages, includingsubject age, weight, gender, diet, and time of administration.

Included in this invention are pharmaceutically acceptable salts andcomplexes of all of the compounds described herein. The salts includebut are not limited to the following acids and bases: Inorganic acidswhich include hydrochloric acid, hydrofluoric acid, hydrobromic acid,hydroiodic acid, sulfuric acid, and boric acid; organic acids whichinclude acetic acid, trifluoroacetic acid, formic acid, oxalic acid,malonic acid, succinic acid, fumaric acid, tartaric acid, maleic acid,citric acid, methanesulfonic acid, trifluoromethanesulfonic acid,benzoic acid, glycolic acid, lactic acid, and mandelic acid; inorganicbases which include ammonia and hydrazine; and organic bases whichinclude methylamine, ethylamine, hydroxyethylamine, propylamine,dimethylamine, diethylamine, trimethylamine, triethylamine,ethyleneamine, hydroxyethylamine, morpholine, piperazine, and guanidine.This invention further provides for the hydrates and polymorphs of allof the compounds described herein.

This invention further provides for metabolites of the compounds of thepresent invention. The in vivo activities and mechanisms of action ofnumerous enzymes responsible for the generation of metabolites ofpharmaceutical compounds are well-known in the art. For example, ethersmay be modified to alcohols, esters may be modified by esterases, oramides may be modified by amidases and peptidases.

This invention further provides a prodrug of the compounds disclosedherein. Knowledge of metabolic activities allows the design of prodrugcompounds which, when administered to a subject, such as a human, areexpected to yield metabolites which include the compounds of the presentinvention.

This invention is additionally directed to a process of making acompound with structure:

which comprises reacting a compound with structure:

with a compound

to form the compound,

wherein Y is

wherein Z is

wherein R1 and R2 (i) are independently H, branched or unbranched C₁-C₆alkyl or alkoxy, branched or unbranched C₂-C₆ alkenyl or alkynyl,branched or unbranched C₁-C₆ hydroxyalkyl, hydroxy, substituted orunsubstituted aryl or aryl-(C₁-C₆)-alkyl, or substituted orunsubstituted heteroaryl or heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, hydroxy, branched orunbranched C₁-C₆ alkyl or alkoxy group, or branched or unbranched C₂-C₆alkenyl or alkynyl group; or (ii) taken together form a substituted orunsubstituted cycloalkyl ring containing 3-10 carbons, wherein thesubstituent if present is a branched or unbranched C₁-C₆ alkyl group orbranched or unbranched C₂-C₆ alkenyl or alkynyl group;

wherein R3 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R4 is H or CH₃;

wherein R5 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂;

wherein R6 is H, branched or unbranched C₁-C₆ alkyl, branched orunbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂; and wherein R14 is independently H or branched orunbranched C₁-C₆ alkyl.

The invention is additionally directed towards a method of treating asubject afflicted with a disease which is susceptible to treatment byantagonism of the human α_(1d) adrenergic receptor which comprisesadministering to the subject an amount of the compound of the inventioneffective to treat the disease.

The treatment of the disease includes the reduction, remission, orregression of the disease or reduction of or relief from symptoms of thedisease.

This invention is directed towards a method of treating a subjectafflicted with hypertension which comprises administering to the subjectan amount of the compound of this invention effective to treathypertension.

This invention is directed towards a method of treating a subjectafflicted with Raynaud's disease which comprises administering to thesubject an amount of the compound of this invention effective to treatRaynaud's disease.

In one embodiment, the compound additionally does not cause hypotensionat dosages effective to treat Raynaud's disease.

This invention is directed towards a method of treating a subjectafflicted with urinary incontinence which comprises administering to thesubject an amount of the compound of this invention effective to treaturinary incontinence.

In one embodiment, the compound additionally does not cause hypotensionat dosages effective to treat urinary incontinence.

This invention is directed towards a method of treating urinaryincontinence in a subject which comprises administering to the subject atherapeutically effective amount of a α_(1d) antagonist which binds tothe human α_(1d) adrenergic receptor with a binding affinity which is atleast ten-fold higher than the binding affinity with which the α_(1d)antagonist binds to (i) a human α_(1a) adrenergic receptor and (ii) ahuman α_(1b) adrenergic receptor, and the α_(1d) antagonist binds to thehuman α_(1d) adrenergic receptor with a binding affinity which isgreater than the binding affinity with which the α_(1d) antagonist bindsto a human 5-HT_(1a) receptor.

In one embodiment, the α_(1d) antagonist binds to the human α_(1d)adrenergic receptor with a binding affinity which is at least 25-foldhigher than the binding affinity with which the α_(1d) antagonist bindsto (i) the human α_(1a) adrenergic receptor and (ii) the human α_(1b)adrenergic receptor, and the α_(1d) antagonist binds to the human α_(1d)adrenergic receptor with a binding affinity which is at least ten-foldhigher than the binding affinity with which the α_(1d) antagonist bindsto the human 5-HT_(1a) receptor.

In one embodiment, the α_(1d) antagonist binds to the human α_(1d)adrenergic receptor with a binding affinity which is at least 25-foldhigher than the binding affinity with which the α_(1d) antagonist bindsto (i) the human α_(1a) adrenergic receptor, (ii) the human α_(1b)adrenergic receptor, and (iii) the human 5-HT_(1a) receptor.

In another embodiment, the α_(1d) antagonist binds to the human α_(1d)adrenergic receptor with a binding affinity which is at least 100-foldhigher than the binding affinity with which the α_(1d) antagonist bindsto (i) the human α_(1a) adrenergic receptor, (ii) the human α_(1b)adrenergic receptor, and (iii) the human 5-HT_(1a) receptor.

In another embodiment, the α_(1d) antagonist additionally does not causehypotension at dosages effective to treat urinary incontinence.

One skilled in the art will readily appreciate that appropriatebiological assays will be used to determine the therapeutic potential ofthe claimed compounds for treating the above noted disorders.

In connection with this invention, a number of cloned human receptorsdiscussed herein, either as plasmids or as stable transfected celllines, have been made pursuant to, and in satisfaction of, the BudapestTreaty on the International Recognition of the Deposit of Microorganismsfor the Purpose of Patent Procedure, and are deposited with the AmericanType Culture Collection, 12301 Parklawn Drive, Rockville, Md. 20852.Specifically, these deposits have been accorded ATCC Accession Numbersas follows in Table 1.

TABLE 1 ATCC Deposits ATCC Cloned Accession Date of Designation ReceptorNo. Deposit Cell line human α_(1d) CRL 11138 Sept. 25, 1992 L-α_(1A)(old human α_(1a)) Cell line human α_(1b) CRL 11139 Sept. 25, 1992L-α_(1B) Cell line human α_(1a) CRL 11140 Sept. 25, 1992 L-α_(1C) (oldhuman α_(1c)) Cell line human 5-HT_(1a) CRL 11889 May 11, 1995 5HT1A-3Cell line human 5-HT_(1Dα) CRL 10421 Apr. 17, 1990 Ltk-8-30-84 (human5-HT_(1D)) Cell line human 5-HT_(1Dβ) CRL 10422 Apr. 17, 1990 Ltk-11(human 5-HT_(1B))

Cell Transfections

Transient transfections of COS-7 cells with various plasmids wereperformed using the DEAE-Dextran method which is well-known to thoseskilled in the art. A plasmid comprising an expression vector for thereceptor of interest was added to monolayers of COS-7 cells bathed in aDEAE-Dextran solution. In order to enhance the efficiency oftransfection, dimethyl sulfoxide was typically also added, according tothe method of Lopata (Lopata, et al., 1984). Cells were then grown undercontrolled conditions and used in experiments after about 72 hours.

Stable cell lines were obtained using means which are well-known in theart. For example, a suitable host cell was typically cotransfected,using the calcium phosphate technique, with a plasmid comprising anexpression vector for the receptor of interest and a plasmid comprisinga gene which allows selection of successfully transfected cells. Cellswere then grown in a controlled environment, and selected for expressionof the receptor of interest. By continuing to grow and select cells,stable cell lines were obtained expressing the receptors described andused herein.

Binding Assays

The binding of a test compound to a receptor of interest was generallyevaluated by competitve binding assays using membrane preparationsderived from cells which expressed the receptor. First, conditions weredetermined which allowed measurement of the specific binding of acompound known to bind to the receptor. Then, the binding of the knowncompound to the receptor in membrane preparations was evaluated in thepresence of several different concentrations of the test compound.Binding of the test compound to the receptor resulted in a reduction inthe amount of the known compound which was bound to the receptor. A testcompound having a high affinity for the receptor of interest woulddisplace a given fraction of the bound known compound at a concentrationlower than the concentration which would be required if the testcompound had a lower affinity for the receptor of interest.

This invention will be better understood from the Experimental Detailswhich follow. However, one skilled in the art will readily appreciatethat the specific methods and results discussed are merely illustrativeof the invention as described more fully in the claims which followthereafter.

Experimental Details

Synthesis

The compounds of Examples 6-20 may be obtained using the methodsdepicted in Schemes 1-5. In the Schemes, m is an integer from 0 to 2; nis an integer from 0 to 2; R1 and R2 (i) are independently H, branchedor unbranched C₁-C₆ alkyl or alkoxy, branched or unbranched C₂-C₆alkenyl or alkynyl, branched or unbranched C₁-C₆ hydroxyalkyl, hydroxy,substituted or unsubstituted aryl or aryl-(C₁-C₆)-alkyl, or substitutedor unsubstituted heteroaryl or heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, hydroxy, branched orunbranched C₁-C₆ alkyl or alkoxy group, or branched or unbranched C₂-C₆alkenyl or alkynyl group; or (ii) taken together form a substituted orunsubstituted cycloalkyl ring containing 3-10 carbons, wherein thesubstituent if present is a branched or unbranched C₁-C₆ alkyl group orbranched or unbranched C₂-C₆ alkenyl or alkynyl group; R3 is H, branchedor unbranched C₁-C₆ alkyl, branched or unbranched C₂-C₆ alkenyl oralkynyl, C₃-C₇ cycloalkyl, C₃-C₇ cycloalkylalkyl, aryl, heteroaryl,aryl-(C₁-C₆)-alkyl, heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl,substituted C₃-C₇ cycloalkyl, substituted aryl, substituted heteroaryl,substituted aryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl,wherein the substituent if present is a halogen, CN, nitro, C₁-C₆ alkyl,OR14, SR14, N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂,or N(R14)CON(R14)₂; R4 is H or CH₃; R5 is H, branched or unbranchedC₁-C₆ alkyl, branched or unbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇cycloalkyl, C₃-C₇ cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂; R6 is H, branched or unbranched C₁-C₆ alkyl, branchedor unbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂; R7 is H, branched or unbranched C₁-C₆ alkyl, branchedor unbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, aryl,aryl-(C₁-C₆)-alkyl, CO₂R14, CON(R14)₂, substituted C₁-C₆ alkyl,substituted aryl, wherein the substituent is N(R14)₂, halogen, OR14 orSR14; R8 is H or CH₃; R9 is H, F, Cl, Br, branched or unbranched C₁-C₆alkyl or alkoxy, CN; R10 is H or F; R11 is H, F, Cl, Br, I, CN, branchedor unbranched C₁-C₆ alkyl or alkoxy; R12 is H, F, Cl, CN, branched orunbranched C₁-C₆ alkyl or alkoxy; R13 is H or F; X is N or CH; with theproviso that when R11 and R12 are each H, then R9 is F; and R14 isindependently H or branched or unbranched C₁-C₆ alkyl.

When the substituents contain (an) amino, amido, carboxylic acid, and/orhydroxyl group(s), it may be necessary to incorporate protection anddeprotection strategies into schemes 1 and 2. Methods forprotection/deprotection of such groups are well-known in the art, andmay be found, for example, in Greene, et al., 1991.

The following examples are merely illustrative of the methods used tosynthesize compounds claimed in this patent. Examples 1-5 describemethods for making compounds that are precursors of the compounds ofthis invention.

Example 1

8-(2-Chloroethyl)-8-azaspiro[4.5]decane-7,9-dione

Compound 1

The synthesis of the compound of example 1 has been described previouslyby others (Wu, Y. H., 1968) and the procedure was modified as follows. Amixture of 3,3-tetramethyleneglutaric anhydride (10.01 g, 59.5 mmol) andethanolamine (7.28 g, 119 mmol) in pyridine (120 mL) was heated atreflux for 3 hours. The solvent was removed and the residue waspartitioned between 1 N HCl (50 mL) and ethyl acetate (50 mL). Theaqueous layer was extracted with ethyl acetate (2×50 mL). The combinedethyl acetate fractions were dried over sodium sulfate and then thesolvent was removed, leaving a clear oil (11.58 g). The oil in benzene(140 mL) and pyridine (7.7 mL) was cooled to 0° C. Thionyl chloride (7.0mL) was slowly added to the mixture and then the solution was heated at60° C. for 60 minutes. The solution was cooled to room temperature andwater (100 mL) was added. The layers were separated and the aqueouslayer was extracted with ethyl acetate (100 mL). The solvent was removedfrom the combined organic fractions and the residue was purified byflash chromatography over silica gel, eluting with hexane/ethyl acetate(3:1). The solvent was removed from fractions with R_(f)=0.3, giving thetitle compound as a pale yellow oil (6.19 g, 26.9 mmol, 45%). ¹H NMR(300 MHz, CDCl₃) δ4.16 (t, 2H, J=6.5 Hz), 3.66 (t, 2H, J=6.3 Hz), 2.63(s, 4H), 1.77-1.67 (m, 4H), 1.55-1.48 (m, 4H).

EXAMPLE 2

8-[(1R)-2-Chloro-1-methylethyl]-8-azaspiro[4.5]decane-7,9-dione

Compound 2

A mixture of 3,3-tetramethyleneglutaric anhydride (1.12 g, 6.66 mmol)and (R)-(−)-2-amino-1-propanol (1.00 g, 13.3 mmol) in pyridine (15 mL)was heated at reflux for 3 hours. The solvent was removed and theresidue was partitioned between 1 N HCl (10 mL) and ethyl acetate (10mL). The aqueous layer was extracted with ethyl acetate (2×10 mL). Thecombined ethyl acetate fractions were dried over sodium sulfate and thenthe solvent was removed, leaving a clear oil (1.92 g). A portion of thisoil (0.70 g) in benzene (9 mL) and pyridine (0.40 mL) was cooled to 0°C. Thionyl chloride (0.40 mL) was slowly added to the mixture and thenthe solution was heated at 60° C. for 90 minutes. The solution wascooled to room temperature and water (10 mL) was added. The layers wereseparated and the aqueous layer was extracted with ethyl acetate (2×10mL). The solvent was removed from the combined organic fractions and theresidue was purified by flash chromatography over silica gel, elutingwith hexane/ethyl acetate (3:1). The solvent was removed from fractionswith R_(f)=0.3, giving the title compound as a pale yellow oil (294 mg,1.21 mmol, 18%). ¹H NMR (300 MHz, CDCl₃) δ5.09-5.04 (m, 1H), 4.17 (t,1H, J=10.5 Hz), 3.66 (dd, 1H, J=11.1, 5.7 Hz), 2.61 (s, 4H), 1.76-1.69(m, 4H), 1.56-1.51 (m, 4H), 1.40 (d, 3H, J=6.9 Hz).

EXAMPLE 3

(2,5-Difluorophenyl)piperazine

Compound 3

A mixture of 2,5-difluoroaniline (2.58 g, 20 mmol) andbis(2-chloroethyl)amine hydrochloride (3.96 g, 22 mmol) in butanol (10mL) was heated at reflux for 24 hours. The mixture was cooled to roomtemperature, sodium carbonate (2.33 g, 22 mmol) was added, and themixture was heated again at reflux. After 2 days, the mixture was cooledto room temperature, hexane (15 mL) and 3 N NaOH (25 mL) were added, andthe resulting layers were separated. The aqueous layer was extractedwith chloroform (3×25 mL) and the combined organic fractions wereflashed over a column of silica gel. The silica gel was further elutedwith a gradient of chloroform to chloroform/methanol (4:1). The solventwas removed from the combined fractions with R_(f)=0.14 [silica gel,chloroform/methanol (4:1)], giving the title compound as a yellow oil(0.606 g, 3.1 mmol, 15%). ESI-MS m/z 199 (MH⁺).

EXAMPLE 4

(2,4,5-Trifluorophenyl)piperazine

Compound 4

A mixture of 2,4,5-trifluoroaniline (2.94 g, 20 mmol) andbis(2-chloroethyl)amine hydrochloride (3.56 g, 20 mmol) in butanol (10mL) was heated at reflux for 24 hours. The mixture was cooled to roomtemperature, sodium carbonate (2.33 g, 22 mmol) was added, and themixture was heated again at reflux. After 2 days, the mixture was cooledto room temperature, hexane (15 mL) and 3 N NaOH (25 mL) were added, andthe resulting layers were separated. The aqueous layer was extractedwith chloroform (3×25 mL) and the combined organic fractions wereflashed over a column of silica gel. The silica gel was further elutedwith a gradient of chloroform to chloroform/methanol (4:1). The solventwas removed from the combined fractions with R_(f)=0.20 [silica gel,chloroform/methanol (4:1)], giving the title compound as a yellow oil(1.028 g, 4.76 mmol, 24%). ESI-MS m/z 217 (MH⁺).

EXAMPLE 5

4-(2,5-Dimethoxyphenyl)piperidine

Compound 5

A solution of tert-butyl lithium in pentane (1.7 M, 1.0 mL)was slowlyadded to a solution of 2-bromo-1,4-dimethoxybenzene (217 mg) intetrahydrofuran (THF) (1.5 mL) at −78° C. The resulting solution wasbrought to room temperature and stirred for 30 minutes. Then thesolution was cooled back down to −78° C. and a solution of1-tert-butoxycarbonyl-4-piperidone (195 mg) was added. The resultingsolution was stirred and allowed to warm to room temperature. After twohours, the reaction was quenched with aqueous NH₄Cl and the solvent wasremoved. The residue was purified by preparative TLC, giving theintermediate alcohol (222.3 mg, 66%). A portion of the alcohol (71.8 mg)was stirred at room temperature in trifluoroacetic acid (TFA) for twohours. The TFA was removed and the residue was purified by preparativeTLC, giving the intermediate tetrahydropyridine (35.4 mg, 76%). Aportion of the tetrahydropyridine (20.8 mg) in methanol (5 mL) washydrogenated (balloon pressure) over 10% palladium on charcoal (5 mg)for 12 hours. The solution was filtered and the solvent was removed,giving the title compound as a white solid (21.0 mg, 100%, 50% overall).ESI-MS m/z 222 (MH⁺).

EXAMPLE 6

8-{2-[4-(2,5-Difluorophenyl)piperazin-1-yl]ethyl}-8-azaspiro[4.5]decane-7,9-dione

Compound 6

A mixture of 1-(2,5-difluorophenyl)piperazine (100 mg, 0.51 mmol) and8-(2-chloroethyl)-8-azaspiro[4.5]decane-7,9-dione (100 mg, 0.44 mmol)was heated with stirring at 160° C. for 5 hours. The residue wasdissolved in methanol, transferred to a preparative thin layerchromatographic plate (silica gel), and eluted with ethyl acetate/hexane(1:1). A band at R_(f)=0.7 was removed and rinsed withchloroform/methanol (4:1). The solvent was removed, giving the titlecompound as a pale yellow oil. ¹H NMR (300 MHz, CDCl₃) δ6.99-6.89(11-line m, 1H), 6.65-6.52 (m, 2H), 3.95 (t, 2H, J=6.5 Hz), 3.03 (t, 4H,J=4.7 Hz), 2.66 (t, 4H, J=4.7 Hz), 2.60 (s, 4H), 2.54 (t, 2H, J=6.6 Hz),1.74-1.69 (m, 4H), 1.55-1.51 (m, 4H); ESI-MS m/z 392 (MH⁺). The titlecompound was dissolved in ether and precipitated by addition of 1N HClin ether, giving a white solid (64.9 mg, 0.15 mmol, 35%): mp 237-239°C., Anal. Calcd. for C₂₁H₂₇N₃F₂O₂·HCl: C, 58.94; H, 6.59; N, 9.82; F,8.88. Found: C, 58.70; H, 6.46; N, 9.68; F, 9.00.

EXAMPLE 7

8-{2-[4-(2,4,5-Trifluorophenyl)piperazin-1-yl]ethyl}-8-azaspiro[4.5]decane-7,9-dione

Compound 7

A mixture of 1-(2,4,5-trifluorophenyl)piperazine (0.94 g, 4.35 mmol) and8-(2-chloroethyl)-8-azaspiro[4.5]decane-7,9-dione (1.00 g, 4.35 mmol)was heated with stirring at 160° C. for 7 hours. The residue waspartitioned between ethyl acetate (40 mL) and saturated aqueous sodiumcarbonate (40 mL). The aqueous layer was extracted with ethyl acetate(2×40 mL) and the combined ethyl acetate fractions dried over sodiumsulfate. The solvent was removed and the residue was purified by flashchromatography over silica gel, eluting with a gradient of hexane tohexane/ethyl acetate (1:1). The solvent was removed from the desiredproduct [R_(f)=0.7, hexane/ethyl acetate (1:1)], leaving a pale tan oilwhich slowly solidified (0.652 g, 1.60 mmol, 37%, mp 230-234° C.). ¹HNMR (300 MHz, CDCl₃) δ6.89 (ddd, 1H, J=11.7, 10.2, 7.5 Hz), 6.74 (dt,1H, J=12.0, 8.1 Hz), 3.95 (t, 2H, J=6.6 Hz), 2.97 (t, 4H, J=4.7 Hz),2.65 (t, 4H, J=4.7 Hz), 2.60 (s, 4H), 2.54 (t, 2H, J=6.5 Hz), 1.74-1.70(m, 4H), 1.55-1.51 (m, 4H); ¹³C NMR (75 MHz, CDCl₃) δ172.7, 151.0 (ddd,J=243.8, 8.5, 1.7 Hz), 146.9 (ddd, J=241.6, 12.2, 3.2 Hz), 144.7 (ddd,J=242.3, 13.9, 12.4 Hz), 137.4 (ddd, J=9.6, 6.1, 2.9 Hz), 107.9 (dd,J=20.7, 4.1 Hz), 106.4 (dd, J=26.6, 21.5 Hz), 55.9, 53.6 (2C), 51.3 (d,2C, J=3.0 Hz), 45.4 (2C), 40.1, 38.0 (2C), 36.9, 24.7 (2C); ESI-MS m/z410 (MH⁺). The title compound was dissolved in ether and precipitated byaddition of 1N HCl in ether, giving a white solid. The solid wasrecrystallized from hot methanol/chloroform (4:1) (with hexane added tocloudiness), giving white flakes (0.43 g, 0.96 mmol, 22%): mp 234-236.5°C., Anal. Calcd. For C₂₁H₂₆N₃F₃O₂·1.2 HCl: C, 55.66; H, 6.05; N, 9.31;F, 12.58. Found: C, 56.06; H, 6.09; N, 9.21; F, 12.20.

EXAMPLE 8

8-{(1R)-2-[4-(2,5-Difluorophenyl)piperazin-1-yl]-1-methylethyl}-8-azaspiro[4.5]decane-7,9-dione

Compound 8

A mixture of 1-(2,5-difluorophenyl)piperazine (20 mg, 0.10 mmol) and(R)-8-(2-chloro-1-methylethyl)-8-azaspiro[4.5]decane-7,9-dione (20 mg,0.082 mmol) was heated with stirring at 160° C. for 5 hours. The residuewas purified by preparative HPLC [reverse phase column (C₁₈ HC),water/methanol (70:30) with 0.1% TFA to water/methanol (40:60) with 0.1%TFA gradient, UV detection at λ254 nm or λ215 nm], giving (after removalof the solvent) the title compound as a pale yellow oil (21.5 mg, 0.05mmol, 66%): ESI-MS m/z 406 (MH⁺).

EXAMPLE 9

8-{(1R)-2-[4-(2,4,5-Trifluorophenyl)piperazin-1-yl]-1-methylethyl}-8-azaspiro[4.5]decane-7,9-dione

Compound 9

A mixture of 1-(2,4,5-trifluorophenyl)piperazine (105 mg, 0.49 mmol) and(R)-8-(2-chloro-1-methylethyl)-8-azaspiro[4.5]decane-7,9-dione (105 mg,0.43 mmol) was heated with stirring at 160° C. for 5 hours. The residuewas dissolved in methanol, transferred to a preparative thin layerchromatographic plate (silica gel), and eluted with ethyl acetate/hexane(1:1). A band at Rf=0.8 was removed and rinsed with chloroform/methanol(4:1). The solvent was removed, giving the title compound as a paleyellow oil (86.2 mg, 0.20 mmol, 47%). ¹H NMR (300 MHz, CDCl₃) δ6.89(ddd, 1H, J=11.7, 10.2, 7.5 Hz), 6.71 (dt, 1H, J=12.0, 8.1 Hz),5.08-4.96 (m, 1H), 3.14 (dd, 1H, J=12.6, 10.5 Hz), 2.92 (t, 4H, J=4.7Hz), 2.73-2.66 (m, 2H), 2.58 (s, 4H), 2.51-2.44 (m, 2H), 2.36 (dd, 1H,J=12.6, 5.4 Hz), 1.75-1.68 (m, 4H), 1.57-1.50 (m, 4H), 1.34 (d, 3H,J=6.9 Hz); ESI-MS m/z 424 (MH⁺). The title compound was dissolved inether and precipitated by addition of 1N HCl in ether, giving a whitesolid: mp 231-235° C.

EXAMPLE 10

8-{(1S)-2-[4-(2,5-Difluorophenyl)piperazin-1-yl]-1-methylethyl}-8-azaspiro[4.5]decane-7,9-dione

Compound 10

A mixture of 1-(2,5-difluorophenyl)piperazine (20 mg, 0.10 mmol) and(S)-8-(2-chloro-1-methylethyl)-8-azaspiro[4.5]decane-7,9-dione (20 mg,0.082 mmol) was heated with stirring at 160° C. for 5 hours. The residuewas purified by preparative HPLC (see Example 8 for conditions), giving(after removal of the solvent) the title compound as a pale yellow oil(10.8 mg, 0.026 mmol, 32%): ESI-MS m/z 406 (MH⁺).

EXAMPLE 11

8-{(1S)-2-[4-(2,4,5-Trifluorophenyl)piperazin-1-yl]-1-methylethyl}-8-azaspiro[4.5]decane-7,9-dione

Compound 11

A mixture of 1-(2,4,5-trifluorophenyl)piperazine (20 mg, 0.093 mmol) and(S)-8-(2-chloro-1-methylethyl)-8-azaspiro[4.5]decane-7,9-dione (20 mg,0.082 mmol) was heated with stirring at 160° C. for 5 hours. The residuewas purified by preparative HPLC (see Example 8 for conditions), giving(after removal of the solvent) the title compound as a pale yellow oil(18.1 mg, 0.043 mmol, 52%): ESI-MS m/z 424 (MH⁺).

EXAMPLE 12

8-{2-[4-(5-Fluoro-2-methoxyphenyl)piperidin-1-yl]ethyl}-8-azaspiro[4.5]decane-7,9-dione

Compound 12

A mixture of 4-(5-fluoro-2-methoxyphenyl)piperidine (20 mg, 0.096 mmol)and 8-(2-chloroethyl)-8-azaspiro[4.5]decane-7,9-dione(20 mg, 0.087 mmol)was heated with stirring at 165° C. for 1 hour. The residue was purifiedby preparative HPLC (see Example 8 for conditions), giving (afterremoval of the solvent) the title compound as a pale yellow oil (2.6 mg,0.0065 mmol, 7%): ESI-MS m/z 403 (MH⁺).

EXAMPLE 13

8-{2-[4-(5-Chloro-2-methylphenyl)piperazino]ethyl}-8-azaspiro[4.5]decane-7,9-dione

Compound 13

A mixture of 1-(5-chloro-2-methylphenyl)piperazine (20 mg, 0.095 mmol)and 8-(2-chloroethyl)-8-azaspiro[4.5]decane-7,9-dione(20 mg, 0.087 mmol)was heated with stirring at 160° C. for 5 hours. The residue waspurified by preparative HPLC (see Example 8 for conditions), giving(after removal of the solvent) the title compound as a pale yellow oil(17.3 mg, 0.043 mmol, 49%): ESI-MS m/z 404 (MH⁺).

EXAMPLE 14

8-{2-[4-(2,6-Difluorophenyl)piperazino]ethyl}-8-azaspiro[4.5]decane-7,9-dione

Compound 14

A mixture of 1-(2,6-difluorophenyl)piperazine (23.8 mg, 0.120 mmol) and8-(2-chloroethyl)-8-azaspiro[4.5]decane-7,9-dione(20 mg, 0.087 mmol) washeated with stirring at 160° C. for 5 hours. The residue was purified bypreparative HPLC (see Example 8 for conditions), giving (after removalof the solvent) the title compound as a pale yellow oil (8.9 mg, 0.023mmol, 26%): ESI-MS m/z 392 (MH⁺).

EXAMPLE 15

8-{2-[4-(3,4-Difluorophenyl)piperazin-1-yl]ethyl}-8-azaspiro[4.5]decane-7,9-dione

Compound 15

A mixture of 1-(3,4-difluorophenyl)piperazine (21.5 mg, 0.109 mmol) and8-(2-chloroethyl)-8-azaspiro[4.5]decane-7,9-dione(20 mg, 0.087 mmol) washeated with stirring at 165° C. for 1 hour. The residue was purified bypreparative HPLC (see Example 8 for conditions), giving (after removalof the solvent) the title compound as a pale yellow oil (13.7 mg, 0.035mmol, 40%): ESI-MS m/z 392 (MH⁺).

EXAMPLE 16

8-{(1R)-1-Phenyl-2-[4-(2,4,5-trifluorophenyl)piperazino]ethyl}-8-azaspiro[4.5]decane-7,9-dione

Compound 16

A mixture of 1-(2,4,5-trifluorophenyl)piperazine (20 mg, 0.093 mmol) and(R)-8-(2-chloro-1-phenylethyl)-8-azaspiro[4.5]decane-7,9-dione (20 mg,0.066 mmol) was heated with stirring at 160° C. for 5 hours. The residuewas purified by preparative TLC, giving the title compound as anoff-white solid (9.5 mg, 0.020 mmol, 30%): ESI-MS m/z 486 (MH⁺).

EXAMPLE 17

8-{(1R)-1-Benzyl-2-[4-(2,4,5-trifluorophenyl)piperazino]ethyl}-8-azaspiro[4.5]decane-7,9-dione

Compound 17

A mixture of 1-(2,4,5-trifluorophenyl)piperazine (20 mg, 0.093 mmol)8-[(1R)-1-benzyl-2-chloroethyl]-8-azaspiro[4.5]decane-7,9-dione (20 mg,0.063 mmol) was heated with stirring at 160° C. for 5 hours. The residuewas purified by preparative TLC, giving the title compound as anoff-white solid (4.7 mg, 0.0094 mmol, 15%): ESI-MS m/z 500 (MH⁺).

EXAMPLE 18 1-(2-Hydroxyethyl)-4-(2-fluorophenyl)piperazine

Compound 18

A mixture of 1-(2-fluorophenyl)piperazine (0.50 g), 2-iodoethanol (0.48g), and potassium carbonate (1.5 g) in dimethylformamide (7 mL) washeated at reflux for three hours. The solvent was removed in vacuo andthe residue was purified by preparative thin-layer chomatography (silicagel, eluting with chloroform/methanol, 9:1), giving the title compoundas a yellow oil (0.41 g, 65%). ¹H NMR (300 MHz, CDCl₃) δ7.09-6.90 (m,4H), 3.67 (t, 2H, J=5.4 Hz), 3.13 (t, 4H, J=4.7 Hz), 2.76 (br s, 1H),2.71 (t, 4H, J=4.8 Hz), 2.63 (t, 4H, J=5.4 Hz). ¹³C NMR (75 MHz, CDCl₃)δ156.3 (d, J=244.1 Hz), 140.6 (d, J=8.8 Hz), 125.1 (d, J=3.4 Hz), 123.2(d, J=8.1 Hz), 119.5 (d, J=2.9 Hz), 116.7 (d, J=20.6 Hz), 60.1, 58.4,53.6 (2C), 51.2 (d, 2C, J=3.0 Hz).

EXAMPLE 19

1-(2-Chloroethyl)-4-(2-fluorophenyl)piperazine

Compound 19

Thionyl chloride (1 mL) was added dropwise to an ice-cooled solution of1-(2-hydroxyethyl)-4-(2-fluorophenyl)piperazine (300 mg) and pyridine (1mL) in benzene (20 mL). After 30 minutes the solution was brought toroom temperature and then the flask was immersed in a 60° C. oil bathfor 1 hour. The layers were separated and the aqueous fraction wasextracted with ethyl acetate (2×20 mL). The combined organic fractionswere dried over sodium sulfate and the solvent was removed in vacuo,giving the desired product as a yellow solid (58.6 mg, 18%). ESI-MS m/z243 (MH⁺).

EXAMPLE 20

8-{2-[4-(2-Fluorophenyl)piperazin-1-yl]ethyl}-8-azaspiro[4.5]decane-7,9-dione

Compound 20

A mixture of 3,3-tetramethylene glutarimide (6.9 mg) and potassiumcarbonate (40 mg) in dimethylformamide (0.5 mL) was stirred at roomtemperature for 30 minutes.1-(2-Chloroethyl)-4-(2-fluorophenyl)piperazine (10.0 mg) was added andthe resulting solution was heated at 120° C. for 3 hours. The solventwas removed and the residue was purified by thin-layer chomatography(silica gel, eluting with hexane/ethyl acetate, 1:1), giving the titlecompound as a white solid (6.2 mg, 40%). ¹H NMR (300 MHz, CDCl₁ ₃)δ7.07-6.87 (m, 4H), 3.96 (t, 2H, J=6.6 Hz), 3.05 (t, 4H, J=4.8 Hz), 2.67(t, 4H, J=4.7 Hz), 2.59 (s, 4H), 2.54 (t, 4H, J=6.6 Hz), 1.73-1.68 (m,4H), 1.55-1.50 (m, 4H). ESI-MS m/z 374 (MH⁺).

Oral Composition

As a specific embodiment of an oral composition of a compound of thisinvention, 100 mg of one of the compounds described herein is formulatedwith sufficient finely divided lactose to provide a total amount of 580to 590 mg to fill a size O hard gel capsule.

Determination of the Binding Affinities of Compounds

Binding affinities were measured for selected compounds of the inventionat three cloned human alpha-1 receptor subtypes, as well as at the5-HT_(1a) receptor.

The binding properties of compounds at the different human receptorswere determined in vitro using cultured cell lines that selectivelyexpress the receptor of interest. These cell lines were prepared bytransfecting the cloned cDNA or cloned genomic DNA or constructscontaining both genomic DNA and cDNA encoding the human receptors asdescribed below.

α_(1d) Human Adrenergic Receptor: The entire coding region. of α_(1d)(1719 bp), including 150 base pairs of 5′ untranslated sequence (5′ UT)and 300 bp of 3′ untranslated sequence (3′ UT), was cloned into theBamHI and ClaI sites of the polylinker-modified eukaryotic expressionvector pCEXV-3, called EXJ.HR. The construct involved the ligation ofpartial overlapping human lymphocyte genomic and hippocampal cDNAclones: 5′ sequence were contained on a 1.2 kb SmaI-XhoI genomicfragment (the vector-derived BamHI site was used for subcloning insteadof the internal insert-derived SmaI site) and 3′ sequences werecontained on a 1.3 kb XhoI-ClaI cDNA fragment (the ClaI site was fromthe vector polylinker). Stable cell lines were obtained bycotransfection with the plasmid α1A/EXJ (expression vector containingthe α_(1a) receptor gene (old nomenclature)) and the plasmid pGCcos3neo(plasmid containing the aminoglycoside transferase gene) into LM(tk-)cells using the calcium phosphate technique. The cells were grown, in acontrolled environment (37° C., 5% CO₂), as monolayers in Dulbecco'smodified Eagle's Medium (GIBCO, Grand Island, N.Y.) containing 25 mMglucose and supplemented with 10% bovine calf serum, 100 units/mlpenicillin g, and 100 μg/ml streptomycin sulfate. Stable clones werethen selected for resistance to the antibiotic G-418 (1 mg/ml), andmembranes were harvested and assayed for their ability to bind[³H]prazosin as described below (see Radioligand Binding assays).

The cell line expressing the human α_(1d) receptor used herein wasdesignated L-α_(1A) (old nomenclature) and was deposited with theAmerican Type Culture Collection, 12301 Parklawn Drive, Rockville, Md.20852, U.S.A. under the provisions of the Budapest Treaty for theInternational Recognition of the Deposit of Microorganisms for thePurposes of Patent Procedure. The cell line expressing the human α_(1d)receptor, was accorded ATCC Accession No. CRL 11138, and was depositedon Sep. 25, 1992.

α_(1b) Human Adrenergic Receptor: The entire coding region of α_(1b)(1563 bp), including 200 base pairs of 5′ untranslated sequence (5′ UT)and 600 bp of 3′ untranslated sequence (3′ UT), was cloned into theEcoRI site of pCEXV-3 eukaryotic expression vector. The constructinvolved ligating the full-length EcoRI brainstem cDNA fragment from λZapII into the expression vector. Stable cell lines were selected asdescribed above. Membranes were harvested and assayed for their abilityto bind [³H]prazosin as described below (see Radioligand Bindingassays). The cell line used herein was designated L-α_(1B) and wasdeposited with the American Type Culture Collection, 12301 ParklawnDrive, Rockville, Md. 20852, U.S.A. under the provisions of the BudapestTreaty for the International Recognition of the Deposit ofMicroorganisms for the Purposes of Patent Procedure. The cell lineL-α_(1B) was accorded ATCC Accession No. CR 11139, on Sep. 29, 1992.

α_(1a) Human Adrenergic Receptor: The entire coding region of α_(1a)(1401 bp), including 400 base pairs of 5′ untranslated sequence (5′ UT)and 200 bp of 3′ untranslated sequence (3′ UT), was cloned into the KpnIsite of the polylinker-modified pCEXV-3-derived eukaryotic expressionvector, EXJ.RH. The construct involved ligating three partialoverlapping fragments: a 5′0.6 kb HincII genomic clone, a central 1.8EcoRI hippocampal cDNA clone, and a 3′0.6 Kb PstI genomic clone. Thehippocampal cDNA fragment overlaps with the 5′ and 3′ genomic clones sothat the HincII and PstI sites at the 5′ and 3′ ends of the cDNA clone,respectively, were utilized for ligation. This full-length clone wascloned into the KpnI site of the expression vector, using the 5′ and 3′KpnI sites of the fragment, derived from vector (i.e., pBluescript) and3′-untranslated sequences, respectively. Stable cell lines were selectedas described above. Membranes were harvested and assayed for theirability to bind [³H]prazosin as described below (see Radioligand Bindingassays). The stable cell line expressing the human α_(1a) receptor usedherein was designated L-α_(1C) (old nomenclature) and was deposited withthe American Type Culture Collection, 12301 Parklawn Drive, Rockville,Md. 20852, U.S.A. under the provisions of the Budapest Treaty for theInternational Recognition of the Deposit of Microorganisms for thePurposes of Patent Procedure. The cell line expressing the human α_(1a)receptor was accorded Accession No. CR 11140, on Sep. 25, 1992.

Radioligand Binding Assays for α₁ Human Adrenergic Receptors:Transfected cells from culture flasks were scraped into 5 ml of 5 mMTris-HCl, 5 mM EDTA, pH 7.5, and lysed by sonication. The cell lysateswere centrifuged at 1000 rpm for 5 min at 4° C., and the supernatant wascentrifuged at 30,000×g for 20 min at 4° C. The pellet was suspended in50 mM Tris-HCl, 1 mM MgCl₂, and 0.1% ascorbic acid at pH 7.5. Binding ofthe α₁, antagonist [³H]prazosin (0.5 nM, specific activity 76.2 Ci/mmol)to membrane preparations of LM(tk-) cells was done in a final volume of0.25 ml and incubated at 37° C. for 1 hour. Nonspecific binding wasdetermined in the presence of 10 μM phentolamine. The reaction wasstopped by filtration through GF/B filters using a cell harvester.Inhibition experiments, routinely consisting of 6 concentrations of thetested compounds, were analyzed using a non-linear regressioncurve-fitting computer program to obtain Ki values.

5-HT_(1a) receptor: The cell line for the 5-HT_(1a) receptor, designated5-HT1A-3, was deposited on May 11, 1995, and accorded ATCC Accession No.CRL 11889. The cDNA corresponding to the ⁵-HT_(1a) receptor open readingframes and variable non-coding 5′- and 3′-regions, was cloned into theeukaryotic expression vector pCEXV-3. These constructs were transfectedtransiently into COS-7 cells by the DEAE-dextran method. Cells wereharvested after 72 hours and lysed by sonication in 5 mM Tris-HCl, 5 mMEDTA, pH 7.5. The cell lysates were centrifuged at 1000 rpm for 5minutes at 4° C., and the supernatant was centrifuged at 30,000×g for 20minutes at 4° C. The pellet was suspended in 50 mM Tris-HCl buffer (pH7.7 at room temperature) containing 10 mM MgSO₄, 0.5 mM EDTA, and 0.1%ascorbate. The affinity of compounds at 5-HT_(1a) receptors weredetermined in equilibrium competition binding assays using[³H]-8-OH-DPAT. Nonspecific binding was defined by the addition of 10 μMmianserin. The bound radioligand was separated by filtration throughGF/B filters using a cell harvester.

5-HT_(1Dα) (5-HT_(1D)) and 5-HT_(1Dβ) (5-HT_(1B)) receptors: The celllysates of the LM(tk-) clonal cell line stably transfected with thegenes encoding each of these 5-HT receptor subtypes were prepared asdescribed above. The cell line for the 5-HT_(1Dα) (5-HT_(1D)) receptor,designated as Ltk-8-30-84, was deposited on Apr. 17, 1990, and accordedATCC Accession No. CRL 10421. The cell line for the 5-HT_(1Dβ)(5-HT_(1B)) receptor, designated as Ltk-11, was deposited on Apr. 17,1990, and accorded ATCC Accession No. CRL 10422. These preparations weresuspended in 50 mM Tris-HCl buffer (pH 7.4 at 37° C.) containing 10 mMMgCl₂, 0.2 mM EDTA, 10 μM pargyline, and 0.1% ascorbate. The affinitiesof compounds were determined in equilibrium competition binding assaysby incubation for 30 minutes at 37° C. in the presence of 5 nM [³H]serotonin. Nonspecific binding was determined in the presence of 10 μMserotonin. The bound radioligand was separated by filtration throughGF/B filters using a cell harvester.

The compounds described above were assayed using cloned human alphaadrenergic receptors and the cloned human serotonin receptors. Thepreferred compounds were found to be selective for the aid receptor. Thebinding affinities of several compounds are illustrated in the followingtable.

TABLE 2 Binding affinities (pKi) of selected compounds of the presentinvention at cloned human α_(1a), α_(1b), α_(1d), and 5-HT_(1a)receptors Compound α_(1a) α_(1b) α_(1d) 5-HT_(1a) Compound 6 6.0 7.3 9.07.6 Compound 7 5.4 6.4 8.7 6.4 Compound 8 5.2 7.2 9.1 7.3 Compound 9 4.86.8 8.9 6.5 Compound 10 5.1 5.4 7.3 5.6 Compound 11 4.9 5.3 6.9 5.5Compound 12 7.1 7.2 9.0 8.1 Compound 13 5.1 5.5 7.4 6.6 Compound 14 5.56.9 8.4 8.1 Compound 15 5.4 5.8 8.1 7.3 Compound 16 5.1 5.3 7.1 Compound17 5.0 5.3 7.1 Compound 20 6.2 7.2 9.1 8.1

In addition, the binding affinities (pKi) of Compound 7 of the presentinvention at the cloned human 5-HT_(1Dα) (5-HT_(1D)) and5-HT_(1Dβ)(5-HT_(1B)) receptors are 6.1 and 5.3, respectively.

Functional Assays

The functional in vitro activity of Compound 7 was characterized inseveral pharmacologically defined α₁ adrenoceptor subtype isolatedtissue assays (Aboud et al., 1993). (Rat epididymal vas deferens wasused to determine α_(1A) functional responses, spleen was used forα_(1B) responses, and thoracic aorta was used for α_(1D) responses.)Experiments were carried out according to the method of Deng et al.,1996, with minor buffer and tissue setup tension modifications. Krebsadditionally contained 0.1 μM desipramine and 10 μM corticosterone toblock neuronal and extra neuronal uptake, respectively, and 0.3 μMidazoxan to eliminate α₂ mediated responses. Aorta and vas deferensstrips were set at 0.5 g resting tension, and the splenic preparationswere tensioned to 1 g. All preparations were equilibrated for 1 hrbefore the addition of drugs.

When added in concentrations up to 10 μM, Compound 7 failed to produceagonist activity at any of the three rat α₁-AR subtype models.Additionally, its antagonist potency was determined by its ability toantagonize phenylephrine (PE) induced contraction in each of thesepreparations. The degree of shift of the PE agonist dose response-curvein the presence of Compound 7 was measured and pK_(B) values werecalculated (mean±s.e.m.). Compound 7 potently inhibited the effects ofPE in the aorta (pK_(B)=8.5±0.2). It was approximately 2000 fold lesspotent in the vas deferens (pK_(B)=5.2±0.2) and approximately 60 foldless effective in the spleen (pK_(B)=6.7±0.2). This functionalcharacterization indicates that Compound 7 is a highly selective, potentantagonist at the α_(1D) adrenergic receptor.

Contribution of α_(1D)-adrenoceptors to Vascular Contractions inElectrically Stimulated vs Non-stimulated Arteries

In order to assess whether α_(1A) and α_(1D)-adrenoceptors aredifferentially innervated in resistance vessels, we compared antagonismby Compound 7 (α_(1D)) and SNAP 6201 (α_(1A)) (Schorn et al.,1999) inrat caudal arteries contracted by transmural stimulation of intrinsicneurons versus norepinephrine (NE) applied directly to the tissue bath.

Experimental Methods

Rings of rat caudal artery were positioned on L-shaped wire holders andsuspended in Krebs' solution for measurement of isometric tension.Propranolol (3 μM), atropine (0.1 μM), and indomethacin (10 μM) wereincluded in the buffer to prevent the activation of β-adrenoceptors andmuscarinic receptors and prostaglandin formation. Tissue tension wasadjusted to 0.5 g and thereafter readjusted twice. Control contractionswere elicited by the application of NE (10 μM) or by electricalstimulation (trains of pulses: rate=0.01 trains/s; train duration=500ms, frequency=9 Hz; pulse duration=4 ms; 90 volts). The effect of theα_(1D)-selective Compound 7 and a selective α_(1A)-adrenoceptorantagonist, each at a concentration 100-fold greater than its K_(B) forthat receptor, was evaluated in terms of: (1) its ability to reverse thecontraction evoked by NE, and (2) its ability to inhibit the contractionevoked by electrical stimulation.

Results

The results of these experiments indicate that the two methods givestrikingly different indications of the role of the α_(1D) adrenoceptorin vascular contraction. When contractions are evoked by the applicationof NE to the tissue bath, a condition in which the agonist has equalaccess to synaptic as well as extrasynaptic receptors, α_(1D) receptoractivation accounts for only 17% of the total contraction (FIG. 2). Incontrast, when contractions are evoked by electrical stimulation ofintrinsic nerves, a situation in which the actions of the released NEare restricted largely to receptors within the neuromuscular synapse,α_(1D) receptor activation accounts for the majority (58%) of the totalcontraction (FIG. 3). These results indicate that the α_(1D)adrenoceptor is the predominant subtype receiving innervation fromsympathetic neurons in resistance arteries.

Effect of α_(1D)-adrenoceptor Blockade in Anesthetized, NormotensiveRats

The potential for α_(1D) selective antagonists as antihypertensiveagents was reported by Deng et al (1996), based on studies with BMY 7378in anesthetized, normotensive rats. BMY 7378 is an antagonist of theα_(1D) adrenoceptor which exhibits marked selectivity over otherα-adrenoceptors. However, BMY 7378 exhibits an even greater affinity at5-HT_(1A) receptors, at which it is a potent partial agonist.Furthermore, activation of central 5-HT_(1A) receptors is known toresult in a reduction in blood pressure. The possibility that thereduction of blood pressure seen with BMY 7378 in urethane-anesthetizedrats was a result of 5-HT_(1A) activation was not considered in thereport by Deng et al. (1996). Therefore, we designed an experiment totest this possibility.

Experimental Methods

Rats were anesthetized with urethane and PE50 cannulae were placed inthe femoral artery and vein for blood pressure monitoring and drugadministration, respectively. After stabilization, rats wereadministered 200 μl saline vehicle or increasing doses of BMY 7378 orCompound 7 at 10 minutes intervals until the reduction in blood pressurereached a plateau. The 5-HT_(1A)-selective antagonist WAY 100635 (1mg/kg) was then administered and blood pressure monitored for anadditional 10 minutes.

Results

FIG. 4 shows that BMY 7378 produces a dose-dependent reduction insystolic, diastolic and mean blood pressure which reaches a maximumeffect at 300 to 1000 μg/kg. The effect of BMY 7378 on each of theseparameters is reversed by the 5-HT_(1A)-selective antagonist WAY 100635,indicating that the effect is mediated predominately through activationof 5-HT_(1A) receptors.

FIG. 5 shows that Compound 7 also reduces blood pressure in anesthetizedrat, although to a lesser extent than BMY 7378. The effect is maximal at100 to 300 μg/kg. In contrast to BMY 7378, however, this effect is notreversed by WAY 100635, indicating that it is solely a result of α_(1D)adrenoceptor blockade.

We conclude therefore that α_(1D) receptor blockade reduces bloodpressure to a lesser extent than was previously thought, and that theeffect is minor in anesthetized, normotensive rats. To further explorethe potential cardiovascular liabilities of α_(1D) adrenoceptorblockade, we conducted experiments in conscious, normotensive rat.

Effect of α_(1D)-adrenoceptor Blockade in Conscious, Normotensive Rats

Experimental Methods

Rats were anesthetized and a PE50 chronic indwelling cannula was placedin the abdominal aorta for measurement of blood pressure. On thefollowing day, 5 rats each were administered vehicle (1% DMSO) orCompound 7 (4.0 mg/kg in 1% DMSO, i.v.) and mean blood pressuremeasurements were recorded at 0, 5, 30, and 60 minutes thereafter.

Results

No statistically significant changes in blood pressure were seenrelative to vehicle-treated animals (FIG. 6).

Discussion of Cardiovascular Results

α_(1D)-adrenoceptor selective antagonists have been postulated to be ofvalue in the treatment of hypertension. In one study (Deng et al., 1996)this conclusion was based on data obtained with the compound BMY 7378 inurethane-anesthetized, normotensive rats. The data presented hereindemonstrate that their data were misinterpreted, i.e. BMY 7378 elicitsits hypotensive effect predominately via activation of 5-HT_(1A)receptors. The more selective α_(1D) antagonist, Compound 7 (see Table2), elicits only a minor reduction in blood pressure in anesthetizedanimals and, more importantly, produces no reduction in blood pressurein conscious animals. Thus, we conclude that selective α_(1D)antagonistswill be devoid of unwanted cardiovascular side effects in normotensiveindividuals.

In addition to the aforementioned studies in normotensive animals, anextensive study of α₁-adrenoceptor subtype involvement has been made inspontaneously hypertensive rats (Scott et al., 1999). These studiesemployed 11 different antagonist with varying selectivity forα₁-adrenoceptor subtypes. These authors concluded that the hypotensivepotency of the antagonists in their study correlated best with theiraffinity for the α_(1D)-adrenoceptor. The data presented in thisapplication, obtained from experiments in normotensive rats, neithersupport nor contradict the results of the Scott et al., 1999 study.

REFERENCES

Aboud, R., et al. (1993) Br. J. Pharmacol. 109:80-87.

Broten, T., et al. (April, 1998) Experimental Biology Meeting, SanFrancisco, Calif., Abstract 2584.

Bylund, D. B. (1992) FASEB J. 6:832-9.

Deng, X. F. et al. (1996) Br. J. Pharmacol. 119:269-276.

Greene, T. W. and Wuts, P. G. M. (1991) Protective Groups in OrganicSynthesis, 2nd Edition John Wiley & Sons, New York.

Hieble, et al. (1995) Pharmacological Reviews 47:267-270.

Lopata, et al. (1984) Nucl. Acids. Res. 12:5707-5717.

McGrath, et al. (1989) Med. Res. Rev., 9:407-533.

Schorn, T., et al. (1999) FASEB J., 13(4), Abstract 150.8.

Scott, A., et al. (1999) FASEB J., 13(4), Abstract 150.6.

Sokoloff, P. et al. (1990) Nature 146:347.

Wu, Y. H. U.S. Pat. No. 3,398,151, 1968.

What is claimed is:
 1. A method of inhibiting activation of a humanα_(1d) adrenergic receptor which comprises contacting the receptor witha compound so as to inhibit activation of the receptor, wherein thecompound has the structure:

wherein each m and n is independently an integer from 0 to 2; whereineach Y and Z is independently

wherein R₁ and R₂ (i) are independently H, branched or unbranched C₁-C₆alkyl or alkoxy, branched or unbranched C₂-C₆ alkenyl or alkynyl,branched or unbranched C₁-C₆ hydroxyalkyl, hydroxy, substituted orunsubstituted aryl or aryl-(C₁-C₆)-alkyl, or substituted orunsubstituted heteroaryl or heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, hydroxy, branched orunbranched C₁-C₆ alkyl or alkoxy group, or branched or unbranched C₂-C₆alkenyl or alkynyl group; or (ii) taken together form a substituted orunsubstituted cycloalkyl ring containing 3-10 carbons, wherein thesubstituent if present is a branched or unbranched C₁-C₆ alkyl group orbranched or unbranched C₂-C₆ alkenyl or alkynyl group; wherein R₃ is H,branched or unbranched C₁-C₆ alkyl, branched or unbranched C₂-C₆ alkenylor alkynyl, C₃-C₇ cycloalkyl, C₃-C₇ cycloalkylalkyl, aryl, heteroaryl,aryl-(C₁-C₆)-alkyl, heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl,substituted C₃-C₇ cycloalkyl, substituted aryl, substituted heteroaryl,substituted aryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl,wherein the substituent if present is a halogen, CN, nitro, C₁-C₆ alkyl,OR₁₄, SR₁₄, N(R₁₄)₂, SO₂N(R₁₄)₂, CO₂R₁₄, SO₃R₁₄, N(R₁₄)COR₁₄, CON(R₁₄)₂,or N(R₁₄)CON(R₁₄)₂; wherein R₄ is H or CH₃; wherein R₅ is H, branched orunbranched C₁-C₆ alkyl, branched or unbranched C₂-C₆ alkenyl or alkynyl,C₃-C₇ cycloalkyl, C₃-C₇ cycloalkylalkyl, aryl, heteroaryl,aryl-(C₁-C₆)-alkyl, heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl,substituted C₃-C₇ cycloalkyl, substituted aryl, substituted heteroaryl,substituted aryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl,wherein the substituent if present is a halogen, CN, nitro, C₁-C₆ alkyl,OR₁₄, SR₁₄, N(R₁₄)₂, SO₂N(R₁₄)₂, CO₂R₁₄, SO₃R₁₄, N(R₁₄)COR₁₄, CON(R₁₄)₂,or N(R₁₄)CON(R₁₄)₂; wherein R₆ is H, branched or unbranched C₁-C₆ alkyl,branched or unbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR₁₄, SR₁₄,N(R₁₄)₂, SO₂N(R₁₄)₂, CO₂R₁₄, SO₃R₁₄, N(R₁₄)COR₁₄, CON(R₁₄)₂, orN(R₁₄)CON(R₁₄)₂; wherein R₇ is H, branched or unbranched C₁-C₆ alkyl,branched or unbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, aryl,aryl-(C₁-C₆)-alkyl, CO₂R₁₄, CON(R₁₄)₂, substituted C₁-C₆ alkyl,substituted aryl, wherein the substituent is N(R₁₄)₂, halogen, OR₁₄ orSR₁₄; wherein R₈ is H or CH₃; wherein R₁₀ is H or F; wherein R₁₁ is H,F, Cl, Br, I, CN, branched or unbranched C₁-C₆ alkyl or alkoxy; whereinR₁₃ is H or F; and wherein R₁₄ is independently H or branched orunbranched C₁-C₆ alkyl.
 2. The method of claim 1, wherein the compoundhas the structure:


3. The method of claim 2, wherein the compound has the structure:


4. The method of claim 3, wherein the compound has the structure:


5. The method of claim 4, wherein the compound has the structure:


6. The method of claim 5, wherein the compound has the structure:


7. A compound having the structure:

wherein each m and n is independently an integer from 0 to 2; whereineach Y and Z is independently

wherein R₁ and R₂ (i) are independently H, branched or unbranched C₁-C₆alkyl or alkoxy, branched or unbranched C₂-C₆ alkenyl or alkynyl,branched or unbranched C₁-C₆ hydroxyalkyl, hydroxy, substituted orunsubstituted aryl or aryl-(C₁-C₆)-alkyl, or substituted orunsubstituted heteroaryl or heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, hydroxy, branched orunbranched C₁-C₆ alkyl or alkoxy group, or branched or unbranched C₂-C₆alkenyl or alkynyl group; or (ii) taken together form a substituted orunsubstituted cycloalkyl ring containing 3-10 carbons, wherein thesubstituent if present is a branched or unbranched C₁-C₆ alkyl group orbranched or unbranched C₂-C₆ alkenyl or alkynyl group; wherein R₃ is H,branched or unbranched C₁-C₆ alkyl, branched or unbranched C₂-C₆ alkenylor alkynyl, C₃-C₇ cycloalkyl, C₃-C₇ cycloalkylalkyl, aryl, heteroaryl,aryl-(C₁-C₆)-alkyl, heteroaryl-(C₁-C₆)-alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR₁₄, SR₁₄,N(R₁₄)₂, SO₂N(R₁₄)₂, CO₂R₁₄, SO₃R₁₄, N(R₁₄)COR₁₄, CON(R₁₄)₂, orN(R₁₄)CON(R₁₄)₂; wherein R₄ is H or CH₃; wherein R₅ is H, branched orunbranched C₁-C₆ alkyl, branched or unbranched C₂-C₆ alkenyl or alkynyl,C₃-C₇ cycloalkyl, C₃-C₇ cycloalkylalkyl, aryl, heteroaryl,aryl-(C₁-C₆)-alkyl, heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl,substituted C₃-C₇ cycloalkyl, substituted aryl, substituted heteroaryl,substituted aryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl,wherein the substituent if present is a halogen, CN, nitro, C₁-C₆ alkyl,OR₁₄, SR₁₄, N(R₁₄)₂, SO₂N(R₁₄)₂, CO₂R₁₄, SO₃R₁₄, N(R₁₄)COR₁₄, CON(R₁₄)₂,or N(R₁₄)CON(R₁₄)₂; wherein R₆ is H, branched or unbranched C₁-C₆ alkyl,branched or unbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR₁₄, SR₁₄,N(R₁₄)₂, SO₂N(R₁₄)₂, CO₂R₁₄, SO₃R₁₄, N(R₁₄)COR₁₄, CON(R₁₄)₂, orN(R₁₄)CON(R₁₄)₂; wherein R₇ is H, branched or unbranched C₁-C₆ alkyl,branched or unbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, aryl,aryl-(C₁-C₆)-alkyl, CO₂R₁₄, CON(R₁₄)₂, substituted C₁-C₆ alkyl,substituted aryl, wherein the substituent is N(R₁₄)2, halogen, OR₁₄ orSR₁₄; wherein R₈ is H or CH₃; wherein R₁₀ is H or F; wherein R₁₁ is H,F, Cl, Br, I, CN, branched or unbranched C₁-C₆ alkyl or alkoxy; whereinR₁₃ is H or F; and wherein R₁₄ is independently H or branched orunbranched C₁-C₆ alkyl.
 8. A compound of claim 7, wherein the compoundhas the structure:


9. A compound of claim 8, wherein the compound has the structure:


10. A compound of claim 9, wherein the compound has the structure:


11. A compound of claim 10, wherein the compound has the structure:


12. A compound of claim 11, wherein the compound has the structure:


13. A pharmaceutical composition comprising a therapeutically effectiveamount of the compound of claim 7 and a pharmaceutically acceptablecarrier.
 14. The pharmaceutical composition of claim 13, wherein theamount of the compound is an amount from about 0.01 mg to about 800 mg.15. The pharmaceutical composition of claim 14, wherein the amount ofthe compound is from about 0.1 mg to about 300 mg.
 16. Thepharmaceutical composition of claim 15, wherein the amount of thecompound is from about 1 mg to about 20 mg.
 17. The pharmaceuticalcomposition of claim 13, wherein the carrier is a liquid.
 18. Thepharmaceutical composition of claim 13, wherein the carrier is a solid.19. The pharmaceutical composition of claim 13, wherein the carrier is agel.
 20. A pharmaceutical composition obtained by combining atherapeutically effective amount of a compound of claim 7 and apharmaceutically acceptable carrier.
 21. A process for making apharmaceutical composition comprising combining a therapeuticallyeffective amount of a compound of claim 7 and a pharmaceuticallyacceptable carrier.
 22. A process of making a compound with structure:

which comprises reacting a compound with structure:

with a compound

to form the compound, wherein Y is

wherein Z is

wherein R1 and R2 (i) are independently H, branched or unbranched C₁-C₆alkyl or alkoxy, branched or unbranched C₂-C₆ alkenyl or alkynyl,branched or unbranched C₁-C₆, hydroxyalkyl, hydroxy, substituted orunsubstituted aryl or aryl-(C₁-C₆)-alkyl, or substituted orunsubstituted heteroaryl or heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, hydroxy, branched orunbranched C₁-C₆ alkyl or alkoxy group, or branched or unbranched C₂-C₆alkenyl or alkynyl group; or (ii) taken together form a substituted orunsubstituted cycloalkyl ring containing 3-10 carbons, wherein thesubstituent if present is a branched or unbranched C₁-C₆ alkyl group orbranched or unbranched C₂-C₆ alkenyl or alkynyl group; wherein R3 is H,branched or unbranched C₁-C₆ alkyl, branched or unbranched C₂-C₆ alkenylor alkynyl, C₃-C₇ cycloalkyl, C₃-C₇ cycloalkylalkyl, aryl, heteroaryl,aryl-(C₁-C₆)-alkyl, heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl,substituted C₃-C₇ cycloalkyl, substituted aryl, substituted heteroaryl,substituted aryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl,wherein the substituent if present is a halogen, CN, nitro, C₁-C₆ alkyl,OR14, SR14, N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂,or N(R14)CON(R14)₂; wherein R4 is H or CH₃; wherein R5 is H, branched orunbranched C₁-C₆ alkyl, branched or unbranched C₂-C₆ alkenyl or alkynyl,C₃-C₇ cycloalkyl, C₃-C₇ cycloalkylalkyl, aryl, heteroaryl,aryl-(C₁-C₆)-alkyl, heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl,substituted C₃-C₇ cycloalkyl, substituted aryl, substituted heteroaryl,substituted aryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl,wherein the substituent if present is a halogen, CN, nitro, C₁-C₆ alkyl,OR14, SR14, N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂,or N(R14)CON(R14)₂; wherein R6 is H, branched or unbranched C₁-C₆ alkyl,branched or unbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂; and wherein R14 is independently H or branched orunbranched C₁-C₆ alkyl.
 23. A method of treating a subject afflictedwith a disease which is susceptible to treatment by antagonism of thehuman α_(1d) adrenergic receptor which comprises administering to thesubject an amount of the compound of claim 7 effective to treat thedisease.
 24. A method of treating a subject afflicted with hypertensionwhich comprises administering to the subject an amount of the compoundof claim 7 effective to treat the disease.
 25. A method of treating asubject afflicted with Raynaud's disease which comprises administeringto the subject an amount of the compound of claim 7 effective to treatthe disease.
 26. A method of claim 25, wherein the compound additionallydoes not cause hypotension at dosages effective to treat Raynaud'sdisease.
 27. A method of treating a subject afflicted with urinaryincontinence which comprises administering to the subject an amount ofthe compound of claim 7 effective to treat the disease.
 28. A method ofclaim 27, wherein the compound additionally does not cause hypotensionat dosages effective to treat urinary incontinence.
 29. A method oftreating urinary incontinence in a subject which comprises administeringto the subject a therapeutically effective amount of a α_(1d) adrenergicreceptor antagonist, which binds to the human α_(1d) adrenergic receptorwith a binding affinity which is at least 10-fold higher than thebinding affinity with which the compound binds to (i) a human α_(1a)adrenergic receptor and (ii) a human α_(1b) adrenergic receptor, and theα_(1d) antagonist binds to the human α_(1d) adrenergic receptor with abinding affinity which is at least ten-fold higher than the bindingaffinity with which the compound binds to a human 5-HT_(1a) receptor,wherein the α_(1d) adrenergic receptor antagonist has the structure:

wherein each m and n is independently an integer from 0 to 2; whereineach Y and Z is independently

wherein R1 and R2 (i) are independently H, branched or unbranched C₁-C₆alkyl or alkoxy, branched or unbranched C₂-C₆ alkenyl or alkynyl,branched or unbranched C₁-C₆ hydroxyalkyl, hydroxy, substituted orunsubstituted aryl or aryl-(C₁-C₆)-alkyl, or unsubstituted heteroaryl orheteroaryl-(C₁-C₆)-alkyl, wherein the substituent if present is ahalogen, CN, nitro, hydroxy, branched or unbranched C₁-C₆ alkyl oralkoxy group, or branched or unbranched C₂-C₆ alkenyl or alkynyl group;or (ii) taken together form a substituted or unsubstituted cycloalkylring containing 3-10 carbons, wherein the substituent if present is abranched or unbranched C₁-C₆ alkyl group or branched or unbranched C₂-C₆alkenyl or alkynyl group; wherein R3 is H, branched or unbranched C₁-C₆alkyl, branched or unbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇cycloalkyl, C₃-C₇ cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂; wherein R4 is H or CH₃; wherein R5 is H, branched orunbranched C₁-C₆ alkyl, branched or unbranched C₂-C₆ alkenyl or alkynyl,C₃-C₇ cycloalkyl, C₃-C₇ cycloalkylalkyl, aryl, heteroaryl,aryl-(C₁-C₆)-alkyl, heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl,substituted C₃-C₇ cycloalkyl, substituted aryl, substituted heteroaryl,substituted aryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl,wherein the substituent if present is a halogen, CN, nitro, C₁-C₆ alkyl,OR14, SR14, N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂,or N(R14)CON(R14)₂; wherein R6 is H, branched or unbranched C₁-C₆ alkyl,branched or unbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, C₃-C₇cycloalkylalkyl, aryl, heteroaryl, aryl-(C₁-C₆)-alkyl,heteroaryl-(C₁-C₆)-alkyl, substituted C₁-C₆ alkyl, substituted C₃-C₇cycloalkyl, substituted aryl, substituted heteroaryl, substitutedaryl-(C₁-C₆)-alkyl, or substituted heteroaryl-(C₁-C₆)-alkyl, wherein thesubstituent if present is a halogen, CN, nitro, C₁-C₆ alkyl, OR14, SR14,N(R14)₂, SO₂N(R14)₂, CO₂R14, SO₃R14, N(R14)COR14, CON(R14)₂, orN(R14)CON(R14)₂; wherein R7 is H, branched or unbranched C₁-C₆ alkyl,branched or unbranched C₂-C₆ alkenyl or alkynyl, C₃-C₇ cycloalkyl, aryl,aryl-(C₁-C₆)-alkyl, CO₂R14, CON(R14)₂, substituted C₁-C₆ alkyl,substituted aryl, wherein the substituent is N(R14)₂, halogen, OR14 orSR14; wherein R8 is H or CH₃; wherein R10 is H or F; wherein R11 is H,F, Cl, Br, I, CN, branched or unbranched C₁-C₆ alkyl or alkoxy; whereinR13 is H or F; and wherein R14 is independently H or branched orunbranched C₁-C₆ alkyl.
 30. The method of claim 29, wherein the α_(1d)antagonist binds to the human α_(1d) adrenergic receptor with a bindingaffinity which is at least 25-fold higher than the binding affinity withwhich the α_(1d) antagonist binds to (i) the human α_(1a) adrenergicreceptor and (ii) the human α_(1b) adrenergic receptor, and the α_(1d)antagonist binds to the human α_(1d) adrenergic receptor with a bindingaffinity which is at least ten-fold higher than the binding affinitywith which the α_(1d) antagonist binds to the human 5-HT_(1a) receptor.31. The method of claim 30, wherein the α_(1d) antagonist binds to thehuman α_(1d) adrenergic receptor with a binding affinity which is atleast 25-fold higher than the binding affinity with which the α_(1d)antagonist binds to (i) the human α_(1a) adrenergic receptor, (ii) thehuman α_(1b) adrenergic receptor, and (iii) the human 5-HT_(1a)receptor.
 32. The method of claim 31, wherein the α_(1d) antagonistbinds to the human α_(1d) adrenergic receptor with a binding affinitywhich is at least 100-fold higher than the binding affinity with whichthe α_(1d) antagonist binds to (i) the human α_(1a) adrenergic receptor,(ii) the human α_(1b) adrenergic receptor, and (iii) the human 5-HT_(1a)receptor.
 33. A method of claim 38, wherein the α_(1d) antagonistadditionally does not cause hypotension at dosages effective to treaturinary incontinence.
 34. The compound of claim 7, wherein the compoundcomprises the (−) enantiomer.
 35. The compound of claim 7, wherein thecompound comprises the (+) enantiomer.
 36. The method of claim 1,wherein the compound binds to the human α_(1d) adrenergic receptor witha binding affinity which is at least 25-fold higher than the bindingaffinity with which the compound binds to (i) the human α_(1a)adrenergic receptor and (ii) the human α_(1b) adrenergic receptor, andthe compound binds to the human α_(1d) adrenergic receptor with abinding affinity which is at least ten-fold higher than the bindingaffinity with which the compound binds to the human 5-HT_(1a) receptor.37. The method of claim 36, wherein the compound binds to the humanα_(1d) adrenergic receptor with a binding affinity which is at least25-fold higher than the binding affinity with which the compound bindsto (i) the human α_(1a) adrenergic receptor, (ii) the human α_(1b)adrenergic receptor, and (iii) the human 5-HT_(1a) receptor.
 38. Themethod of claim 37, wherein the compound binds to the human α_(1d)adrenergic receptor with a binding affinity which is at least 100-foldhigher than the binding affinity with which the compound binds to (i)the human α_(1a) adrenergic receptor, (ii) the human α_(1b) adrenergicreceptor, and (iii) the human 5-HT_(1a) receptor.